Purification and Properties of Bovine Skeletal Muscle Proteasome

Yamamoto, S.; Gerelt, B.; Nishiumi, T.; Suzuki, A.

doi:2005.18.6.884

Article

Animal Products

Asian-Australasian Journal of Animal Sciences 2005;18(6): 884-891.
https://doi.org/10.5713/ajas.2005.884 Published online November 26, 2005.

Purification and Properties of Bovine Skeletal Muscle Proteasome

S. Yamamoto, B. Gerelt, T. Nishiumi, A. Suzuki

Abstract

This paper describes the purification and properties of a multicatalytic proteinase complex, proteasome, from bovine skeletal muscle, in comparision with proteasome prepared from other species or organs. The purified bovine skeletal muscle proteasome exhibited a single band on polyacrylamide gel electrophoresis under nondenaturing conditions. Bovine skeletal muscle proteasome degraded synthetic peptides maximally at pH 8.0. Relative to pH 8.0, activities were gradually decreased with the lowering pH, but the extent of decrease was substrate-dependent, and the activity at pH 5.5 still retained 78-10% of the activity at pH 8.0, indicating the possibility that the proteasome is active in muscle during aging. When the proteasome was heated at 60째C for 15 or 30 min and treated in the presence of 0.0125% SDS, the activity increased over 1.8 and 3.1 times (LLVY (Suc-Leu-Leu-Val-Tyr-NH-Mec) as a substrate), respectively. These results (activation with heat or SDS) indicate that the hydrolytic activity of proteasome was stimulated under mild denaturing conditions. The characteristics of the bovine skeletal muscle proteasome obtained in our experiment were almost the same as those of the proteasome prepared from other species or organs.

Keywords: Bovine; Proteasome; SDS Treatment; Heat Treatment; Immunobloting; Meat Aging